ASPECTS OF THE DISCOVERY

AND DEVELOPMENT OF PLANTDERIVED DRUGS

A. Douglas Kinghorn, Ph.D., D.Sc.
Professor and Jack L. Beal Chair
College of Pharmacy
The Ohio State University

Second FDA/PQRI Conference on Advancing

Product Quality, North Bethesda, MD
(October 5-7, 2015)

OUTLINE OF PRESENTATION

Rationale for the Search for New

Drugs from Higher Plants.
General
Approaches
to
Drug
Discovery from Tropical Plants in an
Academic Environment.
Development of Silvestrol from Aglaia
foveolata (a Potential Anticancer
Agent) and Pentalinonsterol from
Pentalinon andrieuxii (a Potential
Antileishmanial Agent).
Summary and Conclusions.

KEY QUESTIONS TO BE
ANSWERED
In natural product drug discovery
screening programs in an academic
environment, how, why, and when
may highly promising compounds
be identified?
What
types
of
collaborative
investigations can then be pursued
in order to develop these lead
compounds further?

RATIONALE FOR THE SEARCH

FOR NEW DRUGS FROM HIGHER
PLANTS

SMALL-MOLECULE NATURAL PRODUCT BASED

DRUGS (JANUARY 1, 1981 SEPTEMBER 9, 2012)

Total number of small molecule approved drugs world-wide was 1115

N_all refers to sum of N, NB and ND codes
S*_all refers to sum of S* and S*/NM codes
(Newman and Cragg 2012)

NATURAL PRODUCT-DERIVED
DRUGS INTRODUCED 2000-2008
Source Organism Type
Terrestrial Plants
(apomorphine HCl, arteether, dronabinol/cannabidiol
(mixture), galanthamine HBr, lisdexamfetamine,
methylnatrexone Br, nitisinone, tiotropium Br)
Terrestrial Microorganisms

Number
8

24

(amrubicin HCl, anidulafungin, biapenem, caspofungin acetate,

cefditoren pivoxil, ceftobiprole medocaril, daptomycin, doripenem,
ertapenem, everolimus, fumagillin, gentumazumab ozogamicin,
ixabepilone, micafungin Na, miglustat, mycophenolate Na,
pimecrolimus,
pitavastatin,
retapamulin,
rosuvastatin
Ca,
telithromycin, temsirolimus, tigecycline, zotarolimus)

Marine Organism

(trabectidin, ziconotide)

Terrestrial Animals

(bivalirudin; exenatide; synthetic versions of natural forms)

[Chin et al., AAPS J., 8 (2), E239 (Article 28), 2006; http://www.aapsj.org; Butler, in Natural Product
Chemistry for Drug Discovery, eds. A.D. Buss and M.S. Butler, RSC: Cambridge, U.K., 2010; p. 321]

PLANT NATURAL PRODUCTS AND DERIVATIVES

APPROVED BY THE U.S. FDA (2012 TO MID-2013)
Year

Generic Name

2012 Ingenol mebutate

Lorcaserin
hydrochloride
Omacetaxine
2012
mepesuccinate
2012

2012 Crofelemer

Natural Lead
Compound
Ingenol-3angelate

aAlthough

Indication

Picato

Actinic keratosis

Ephedrine

Belviq

Obesity

Homoharringtonine

Synribo

Croton lechleri
oligomeric proanthocyanidins

Fulyzaq

ado-Trastuzumab
2013 emtansine (protein- Maytansinea
bound)
2013 Ospemifene

Trade Name

Phytoestrogens

Chronic myeloid
leukemia
HIV/AIDS antiretroviralassociated
diarrhea

Kadcyla

Breast cancer

Osphena

Menopauseassociated
dyspareunia

first isolated from a plant, maytansine is now regarded as a microbial product.

GENERAL APPROACHES TO
COLLABORATIVE DRUG DISCOVERY
FROM TROPICAL PLANTS IN AN
ACADEMIC ENVIRONMENT

PLANT NATURAL PRODUCT DRUG DISCOVERY

AND DEVELOPMENT
Natural Products Discovery
Medicinal Chemistry
Molecular Modeling
Target-based Bioassays
Cell-based Bioassays
In Vivo Bioassays

Lead Identification

Medicinal Chemistry
Combinatorial Chemistry

Lead Optimization

Pharmacology, Toxicology
Pharmacokinetics, ADME
Drug Delivery

Lead Development

Drug Candidates

Clinical Trials

MAJOR STAGES IN NATURAL

PRODUCT DRUG DISCOVERY
Organism collection (after development of intellectual property
agreements).
Preparation of extracts (using standardized extraction scheme).
Initial bioassays (cell-based and target-based).
Biostatistics; data management; dereplication of leads; lead
prioritization).
Bioactivity-directed fractionation (= isolation of active
compounds from biomass using a decision tree based solely on
bioactivity).
Structure elucidation of bioactive compounds.
Scale up and analogue development of lead compounds.
Advanced bioassays; data management, biostatistics).
Lead optimization; pharmaceutical development.
(Clark, In Foyes Principles of Medicinal Chemisry, 5th Edn., Williams, D.A.;
Lemke, T.L., Eds., Lippincott Williams & Wilkins: Baltimore, 2002, p. 24)

CONVENTION ON
BIOLOGICAL DIVERSITY (CBD)

A treaty with 42 Articles dictating codes of behavior

in the study and sustainable use of biological
diversity (http://www.biodiv.org/).
This was signed by >150 countries during the 1992
Earth Summit in Rio de Janeiro (Rio Convention).
The U.S.A. signed in 1993, but never ratified this
treaty. In practice, this means the government must
follow the treaty, but there is no binding effect on
private citizens.
Source countries have sovereign right over their
genetic resources (e.g., mutually agreed terms, prior
informed consent, equitable sharing of benefits).

(Cordell, in Natural Product Chemistry for Drug Discovery, eds. A.D.

Buss and M.S. Butler, RSC: Cambridge, U.K., 2010; p. 81)

NUMBERS OF ORGANISMS
FOR DRUG DISCOVERY
Eubacteria (bacteria), cyanobacteria (blue-green algae)
4,000a
Archaea (halobacteria, cyanogens)
Protoctista (e.g., protozoa, diatoms, algae, including red algae
and green algae)

80,000

Plantae (mosses and liverworts, ferns, seed plants)b

270,000

Fungi (e.g., molds, lichens, yeasts, mushrooms)c

Animalia (e.g., mesozoa, sponges, jellyfish, corals, flatworms,
roundworms, sea urchins, mollusks, segmented worms,
arthropods, insects, fish, amphibians, birds, mammals)d-f
a

72,000
1,320,000

Figures are species described taxonomically to date in each group.

Plants are the second largest group of classified organisms, representing 15% of the known biodiversity.
c Only a relatively small proportion (5%) of the estimated 1.5 m fungi have been classified taxonomically to date.
d The largest numbers of organisms are the arthropods, inclusive of insects (ca. 950,000 species).
e Of the 28 major animal phyla, 26 are found in a marine environment.
f Over 200,000 species of invertebrate animals and algal species occur in the sea.
b

(Tan et al., Curr. Drug Targets 7, 265, 2006)

SECONDARY METABOLITES FROM VASCULAR

PLANTS (TRACHEOPHYTES; HIGHER PLANTS)
Altogether there are an estimated 200,000 secondary
metabolites (natural products) (Dixon and Strack,
Phytochemistry 62, 815, 2003).
Of these, the major groups are estimated as
isoprenoids (ca. 80,000 compounds), phenolics (ca.
40,000 compounds), and alkaloids (ca. 30,000
compounds).
More secondary metabolites have been isolated from
plants than other types of organisms.
An average leaf may contain >30,000 phytochemicals
(Turi et al., J. Nat. Prod. 78, 953, 2015).
Specialized defensive secondary metabolites are
associated with higher plants, such as gallotannins,
proanthocyanidins, and resveratrol oligomers.

NUMBER OF NEW NATURAL PRODUCT

COMPOUNDS PUBLISHED IN THE
JOURNAL OF NATURAL PRODUCTS
OVER THE DECADE 2005-2014a,b
Terrestrial
Microbes/
Fungi

Terrestrial
Plantsc

Terrestrial
Animals

Marine /
Aquaticd

Otherse

1,869
(14.5%)

7,760
(60.0%)

117
(0.9%)

3,138
(24.3%)

33
(0.3%)

From a total of 12,917 new small-molecule compounds. Percentages

of the total are shown in parentheses.
b Annual totals of new compounds reported were: 2005, 1,200; 2006,
1,276; 2007, 1,269; 2008, 1,388; 2009, 1,505; 2010, 1,369; 2011, 1,303;
2012, 1,049; 2013, 1,156; 2014, 1,402.
c Including higher and lower plants (vascular and non-vascular).
d Including animals, microorganisms, and plants.
e For example, propolis of different geographical origins.
a

DEVELOPMENT OF SILVESTROL
(A POTENTIAL ANTICANCER AGENT)
AND PENTALINONSTEROL
(A POTENTIAL ANTILEISHMANIAL
AGENT)

DRS. M.E. WALL (LATE) AND M.C. WANI:

CO-DISCOVERERS OF TAXOL AND
CAMPOTHECIN

(Photograph by Jimmy W. Crawford, RTI)

COLLABORATIVE PROJECTS ON THE DISCOVERY

OF NATURAL PRODUCT ANTICANCER AGENTS
National Cooperative Drug Discovery Group (NCDDG) Granta
(U19 CA52956) (1990-1995; 1995-2000; 2000-2006)
Program Project Grantb (P01 CA125066) (2007-2013; 2014-2019)

Current Collaboration is between:

The Ohio State University (OSU), Columbus, OH;
University of Illinois at Chicago (UIC), Chicago, IL;
University of North Carolina at Greensboro (UNC-G), NC;
Mycosynthetix Inc., NC;
Eisai Inc., Andover, MA
Both grants funded by the United States National Cancer
Institute, NIH
a,bPrincipal Investigators: G.A.Cordell (UIC; 1990-1992);
A.D. Kinghorn (UIC/OSU; 1992-present)
[Most recent review: Kinghorn et al., Pure Appl. Chem. 81, 1051, 2009]

PLANT COLLECTIONS AT UIC (DRS. NORMAN

FARNSWORTH AND DOEL SOEJARTO)

Some tropical forests support more tree species in 0.5 km2

than in all of North America or Europe (Burslem, 2001).

About 120,000 endemic species in tropical areas regarded
as threatened due to massive habitat loss (Pitman &
Jrgensen, 2002).
Source countries have sovereign right over their genetic
resources (e.g., mutually agreed terms, prior informed
consent, equitable sharing of benefits).

Anticancer Plant Collections 1990-2011

Total plant accessions obtained 6,599
Representing:
2,609 species
1,466 genera
222 families
Over 200 recollections

FORMER RAID PROGRAM OF THE UNITED

STATES NATIONAL CANCER INSTITUTE
Rapid Access to Intervention Development

(http://dtp.nci.nih.gov/docs/raid/raid_index.ht
ml).
A program designed to facilitate translation to

the clinic of novel, scientifically meritorious

therapeutic interventions originating in the
academic community.
Contracts leading to preclinical development

leading to filing of an IND.

Required submission of a formal proposal

and peer review.

BETULINIC ACID: A NCDDG COMPOUND THAT

ENTERED CLINICAL TRIALS AT THE
UNIVERSITY OF ILLINOIS AT CHICAGO

H
H

COOH

H
HO

Betulinic acid was isolated from Ziziphus mauritania as a selective, nontoxic antimelanoma agent (Pisha et al., Nature Med. 1, 1046, 1995).
Preclinical development of betulinic acid was supported through the
RAID program of NCI (cycles III and VI, T.K. Das Gupta, Principal
Investigator).
Has entered Phase I/II clinical trials as a 20% ointment for topical
treatment of dysplastic nevus syndrome with the potential to transform
to melanoma (http://clinicaltrials.gov/; NCT00346502).

SCHEME OF ORGANIZATION FOR

PROGRAM PROJECT 2P01 CA125066-07
EXTERNAL ADVISORY COMMITTEE
William H. Gerwick (Scripps Institute of Oceanography, San Diego)
Susan B. Horwitz (Yeshiva University)
George R. Pettit (Arizona State University)
William C. Rose (formerly Bristol-Myers Squibb, Princeton, N.J.)
PROJECT 1 The Ohio
State University (OSU)
Tropical Plants,
Biological Testing
A.D. Kinghorn, PL
E.J. Carcache de Blanco
D.M. Lucas

CORE A - UIC
Biological Testing
J.E. Burdette, CD*
S.M. Swanson
(University of Wisconsin)

PROJECT 2 University of
Illinois at Chicago (UIC)
Aquatic Cyanobacteria
Plant Collections
J. Orjala, PL
D.D. Soejarto

PROJECT 3 University of North

Carolina Greensboro (UNC-G)
Filamentous Fungi, Biological Testing

CORE B OSU
Medicinal Chemistry;
Pharmacokinetics
J.R. Fuchs, CD
M.A. Phelps
NCI PROJECT OFFICER
Yali Fu

N.H. Oberlies, PL
C. J. Pearce (Mycosynthetix Inc.)
B.R. Stockwell (Columbia University)
M.C. Wani (Consultant) (RTI)

CORE C - OSU
Administration/Biostatistics
A.D. Kinghorn, CD
L.-H. Xu, X. Zhang
Y. Shen (Eisai Inc.)

NExT PROGRAM OF THE UNITED STATES

NATIONAL CANCER INSTITUTE (NCI)
The NCI Experimental Therapeutics (NExT) Program

is designed to enable and foster the clinical

translation and successful commercialization of novel
therapeutic interventions, either synthetic, natural
product, or biologic, arising from academic, private or
governmental entities (http://next.cancer/gov).
Proposals for drug discovery and development
resources and expertise are examined by an
extramural panel.
The panel reviews the applications for scientific merit
and feasibility, novelty, and clinical need.
Proposals may be selected for entry into various
points in the NeXT pipeline (e.g., lead development,
preclinical toxicology, and phase 0/I clinical trials).

STRUCTURAL CHARACTERIZATION OF
SILVESTROL FROM AGLAIA FOVEOLATA
HO

H
OH

OCH3

OCH3
OH OH
COOCH3
O

Silvestrol

OCH3

X-ray Structure by
Drs.
Bernard
Santarsiero and
Andrew Mesecar
(UIC)
Drs. Bang Yeon Hwang and Baoning Su

(Hwang et al., J. Org. Chem. 69,

3350, 2004; ibid., 69, 6156)

AGLAIA FOVEOLATA (MELIACEAE)

COLLECTED IN INDONESIA

Aglaia foveolata Pannell (Meliaceae), is a tree growing in lowland

forests in Borneo (Brunei, Indonesia, Malaysia), with edible fruits.
This plant (fruits and twigs) was collected initially in Kalimantan,
Indonesia in 2000, but misidentified as Aglaia silvestris (M.
Rohmer) Merrill.
The voucher specimen for the original collection was re-identified
by Dr. Christine Pannell, Daubeny Herbarium, University of Oxford
as Aglaia foveolata Pannell (Hwang et al., J. Org. Chem. 69, 6956,
2004).

IN VITRO CYTOTOXICITY OF SILVESTROL

Compound
Silvestrol
Methyl
rocaglateb
Paclitaxelc
Camptothecinc
a

Lu1
1.2
163
2.3
28.7

Cell linea
LNCaP
MCF-7
1.5
1.5
325
Not
determined
4.7
28.7

0.7
28.7

HUVEC
4.6
203
105.5
258.6

ED50 values (nM) Lu1 = human lung cancer; LNCaP =

hormone-dependent human prostate cancer; MCF-7 =
human breast cancer; HUVEC = human umbilical vein
endothelial cells.
Isolated in previous work from Aglaia rubiginosa (RiveroCruz et al., J. Nat. Prod., 67, 343, 2004).
Used as positive control.

(Hwang et al., J. Org. Chem. 69, 3350, 2004)

EFFECT OF SILVESTROL IN THE IN VIVO HOLLOW

FIBER TEST AND IN THE MURINE P-388 LEUKEMIA
MODEL
Hollow fiber:
Murine P-388 leukemia:
Active at maximum tolerated dose of
2.5 mg/kg/inj, given by intraperitoneal
injection daily for five consecutive
days (qd5) in ip P388 model.
Achieved maximum lifespan increase
corresponding to T/C of 150%.
Inactive (T/C = 100%) in iv P388
leukemia model when administered
by either the iv or ip route using a
daily times five schedule (qd5).
Active (T/C = 129%) in same tumor
model when injected iv on a twicedaily schedule (2qd5) using the
same cumulative dose (2 mg/kg/day).

(Hwang et al., J. Org. Chem. 69,

3350, 2004)

ISOLATION OF SILVESTROL FROM A RECOLLECTION

OF AGLAIA FOVEOLATA FROM INDONESIA
Plant Material
Extract with MeOH
An aliquot (500 g, 5% w/w of
the total dried methanol
extract) was used for work-up

MeOH Extract

A 40-45 kg recollection of
Aglaia foveolata stem bark
was collected in the Fall of
2007, in Kalimantan

Dissolve in mixture of MeOH-H2O (9:1)

Defat with hexanes

Hexanes Extract

Residue
Add H2O, partition with CHCl3

This extract was

active against the
HT-29 cell line
(IC50 = 0.4 g/ml)

Aqueous Extract

CHCl3 Extract
Wash with 1% NaCl

Partially Detannified CHCl3 Extract

Subjected to separation over a Si gel column
(CH2Cl2acetone, 20:1 to 100% acetone)

Subfractions F01-F08

Chromatography of active fraction

F07 (7 g; IC50 = <0.016 g/ml) led to
the purification of silvestrol (100 mg)

Large-scale
column
chromatography of a
recollection of Aglaia
foveolata,
from
Indonesia, leading to
purified
silvestrol
(conducted for the
United States National
Cancer Institute)

Dr. Thomas McCloud,

SAIC-Frederick, MD

% Live Cells Relative to Untreated

EFFECTS OF SILVESTROL ON B AND T

CELLS IN WHOLE BLOOD FROM CLL
PATIENTS
B Cells

T Cells

120

120

100

100

80

80

60

60

40

40

20

20

24
48
72
Incubation Time (hours)

80 nM Silvestrol

24

48

72

Incubation Time (hours)

1 M 2-F-ara A

(Lucas et al., Blood 113, 4656, 2009)

SILVESTROL SIGNIFICANTLY IMPROVED

SURVIVAL IN AN ACUTE LYMPHOBLASTIC
LEUKEMIA XENOGRAFT MOUSE MODEL

% Survival

100

Median survival
difference P = 0.002

80

Control (N=13)

60

Silvestrol (N=14)

40
20

1.5 mg/kg i.p. M, W, F; started 1

wk post-engraftment

Treatment stopped

0
0

10

15

20

25

30

35

40

45

85

Days Post Engraftment

The remaining three mice appeared normal 12+ weeks postengraftment, 6 weeks after the last treatment.
(Lucas et al., Blood 113, 4656, 2009)

SILVESTROL: ACTIVITY AS A TRANSLATION

INHIBITOR

Pelletier and co-workers have shown that silvestrol is a translation

inhibitor by targeting eukaryotic initiation factor (eIF) 4A (Cencic et
al., PLoS ONE 4(4), e5223, 2009).
In recent work, using biotinylated 5-epi-silvestrol, a specific
interaction was shown with eIF4AI and eIF4AII (Chambers et al.,
Org. Lett. 15, 1406, 2013).

(Lucas et al., Curr. Drug Targets 11, 811, 2010)

SILVESTROL: A POTENTIAL NEW THERAPEUTIC

AGENT FOR B-CELL MALIGNANCIES
O
HO

O
OH

OMe

OMe
OH OH

COOMe
O

OMe

At The Ohio State University, silvestrol has shown promising in

vivo activity in models of acute lymphoblastic leukemia, acute
myeloid leukemia, EBV-driven lymphoma, and mantle cell
lymphoma as well as interesting immunomodulatory activity
(Lucas et al., Blood 113, 4656, 2009; Alinari et al., Clin. Cancer
Res. 18, 4600, 2012; Alahkar et al., J. Hematol. Oncol. 6, 21, 2013;
Patton et al., Oncotarget 6, 2693, 2015).
The mechanism of antileukemic action was further investigated
through a NCI/NIH SPORE (P50) award (Dr. J. C. Byrd, PI) (work
by Drs. M.R. Grever and D.M. Lucas; 2009-2015).
Silvestrol has been undergoing preclinical toxicology
development through the NCI NExT pipeline (P.I., M.R. Grever).

AGREEMENT FOR DEVELOPMENT OF SILVESTROL

FOR POTENTIAL TREATMENT OF B-CELL MALIGNANCIES
In June 2012, an agreement was
signed
to
jointly
develop
silvestrol between The Ohio State
University and the Sarawak
Biodiversity Center, with the
immediate goal of conducting
preclinical toxicology.
This slide shows the two other
faculty participants in this work
at Ohio State, Drs. Michael
Grever (top left) and David Lucas
(top
right),
and
Dr.
Rita
Manurung,
former
Chief
Operating
Officer,
Sarawak
Biodiversity Center, Kuching,
Sarawak, Malaysia (bottom).
Silvestrol may be sourced from
Aglaia
stellatopilosa
Pannell
obtained from Sarawak.

ATTRIBUTES OF SILVESTROL AS A CANDIDATE

ANTICANCER AND IMMUNOMODULATORY COMPOUND

The compound is a new composition of matter

representing a relatively unstudied structural type.
Silvestrol has shown efficacy at non-toxic doses in a
range of murine in vivo models germane to human
cancer, including some rare diseases that are difficult to
treat. It acts differentially on different immune subsets.
The presently known mechanism of action of the
compound is unusual (an inhibitor of protein translation,
acting specifically at eukaryotic initiation factor eIF4AI/II).
Sensitizes tumors to other agents, so its use in
combination therapy may be feasible.
Silvestrol has overall favorable pharmacokinetics when
injected ip or iv in mice, and may be formulated in
hydroxypropyl--cyclodextrin.
Intellectual property (IP) protection has been established.
A reliable supply of the compound is available (obtained
from Aglaia stellatopilosa grown in Sarawak, Malaysia).

ACKNOWLEDGMENTS
(STUDIES ON PLANT ANTICANCER AGENTS)
Chemistry

Biological Testing

Prof. Geoffrey A. Cordell (University of

Illinois at Chicago, UIC)
Prof. James R. Fuchs (The Ohio State
University, OSU)
Prof. Andrew D. Mesecar (UIC)
Prof. Bernard D. Santarsiero (UIC)
Dr. David J. Newman (NCI-Frederick)
Dr. Thomas G. McCloud (SAIC-Frederick)
Dr. Bang Yang Hwang (UIC)
Dr. Li Pan (OSU)
Dr. Angela A. Salim [(OSU)]
Dr. Baoning Su (UIC; OSU)

Dr. Hee-byung Chai (UIC; OSU)

Dr. Robert A. Baiocchi (OSU)
Dr. Stuart Emanuel [Bristol-Myers
Squibb (B-MS)]
Dr. Craig R. Fairchild [BristolMyers Squibb (B-MS)]
Prof. Michael R. Grever (OSU)
Dr. David M. Lucas (OSU)
Prof. John M. Pezzuto (UIC)
Dr. William C. Rose (B-MS)
Prof. Steven M. Swanson (UIC)
Dr. Robert Wild (B-MS)

Plant Collection/Taxonomy
Prof. Norman R. Farnsworth (UIC)
Prof. Djaja D. Soejarto (UIC)
Dr. Caroline M. Pannell (University of
Oxford)
Dr. Leonardus B.S. Kardono (Indonesia)
Dr. Soedarsono Riswan (Indonesia)

Funding obtained from NIH/NCI

CA52956 and P01 CA125066 (to A.D.
Kinghorn) is gratefully acknowledged

BACKGROUND TO LEISHMANIASIS
This is a protozoan vector borne parasitic disease (transmitted
by a sand fly).
About 12 million people infected worldwide, with two million
new cases annually, and 350 million people at risk.
Severe public health problem in parts of East Africa and the
Middle East, the Indian subcontinent, and Central and South
America.
Existing drug therapy has several drawbacks including the
development of resistance, toxicity, and poor compliance.

http://www.who.int/leishmaniasis/disease_
epidemiology/en/index.html

APPROACH TO NOVEL DRUG

DISCOVERY FOR LEISHMANIASIS
Screening of plant materials from Mexico.
Isolation and characterization of bioactive constituents.
Development of efficient synthetic methods to prepare the active
compounds and derivatives for MOA, SAR, and optimization
studies.
United States

Mexico

Riparian forest near

Hopelchen, Campeche
(Mexico)
C. M. Lezama-Dvila and
A. P. Isaac-Mrquez

http://www.ecoyuc.com/

SCREENING OF PENTALINON ANDRIEUXII

ROOTS FOR ANTILEISHMANIAL ACTIVITY

Traditionally in Campeche, Mexico,

skin
lesions
from
cutaneous
leishmaniasis are treated topically by
Mayan healers with an infusion of the
roots of Pentalinon andrieuxii B.F.
Hansen & Wunderlin (Apocynaceae).
The inner part of the roots of is then
tightly fixed to the skin lesions. This
procedure is repeated daily until the
wound heals.
In a preliminary laboratory study, in
vitro inhibitory activity against
Leishmania mexicana amastigotes
was found for the root hexane extract
of P. andreuxii (PARE).
(Lezama-Davilla et al., Fitoterapia 78, 255,
2007)

Air-dried Roots of P. andrieuxii (by Y. Deng)

Aerial Parts of P. andrieuxii

www.nybg.org/.../Pentalinon_andrieuxii.html

EFFECT OF PENTALINON ANDRIEUXII

ROOT HEXANE EXTRACT (PARE) ON THE
LEISHMANIA MEXICANA PARASITE IN VIVO
A preliminary in vivo experiment with the root extract was performed using
10 week old male C57BL/6 mice, which were infected with L. mexicana
promastigotes in the ear dermis.

Mice were treated with 10 g of PARE dissolved in 50 l of DMSO/PBS. The extract was applied on
the infected ear daily for 21 days [note the lymphocytes and macrophages (white cells)].

(Lezama-Davilla et al., Phytother. Res. 20, 909, 2014)

ANTILEISHMANIAL STEROLS ISOLATED

FROM PENTALINON ANDRIEUXII ROOTS
R

17

H
H

H
H

1 R=

H
HO

HO

H
O

8 R=

4 R=

9 R=

O
3 R=

17

13 R =

7 R=

14 R =

18 R = OCH3
19 R = H
OH

O
COOH

H
5 R=

6 R=

10 R =

H
O

H
AcO

15

11 R =

OH

20
H 20

21

H
H

Active compound
New compound

12 R =

H3CO

OH

16

(Li et al., Phytochemistry 82, 128, 2012)

O O
2

O
O

O
H

SYNTHESIS OF PENTALINONSTEROL
(1) FROM PREGNENOLONE (2)
1. TBSCl,
imidazole,
DMF
95%

2. LDA, HMPA,
THF, -78C;
HO

Br

1. Tebbe reagent
toluene, THF
50%
2. TBAF, THF
94%

78%

21
20

TBSO

22

R
17

Al(OiPr)3,
N-Me piperidone
3

pentalinonsterol

89%

HO

R=

Drs. James R. Fuchs and Dalia Abdelhamid

(The Ohio State University)

(Gupta et al., ACS Inf. Dis., 2015, under revision)

ADDITIONAL WORK PERFORMED ON

PENTALINONSTEROL

Pentalinonsterol (>30 mg) was synthesized from pregnenolone

in five synthetic steps.
In an in vivo BALB/c mouse model of visceral leishmaniasis
induced by Leishmania donovani, i.v. treatment with liposomal
encapsulated pentalinonsterol (2.5 mg/kg) led to a significant
reduction in parasite burden in the in the liver and spleen.
Infected mice showed a strong host-protective TH1 immune
response with IFN- production and the formation of matured
hepatic granulomas, when treated with the liposomal
encapsulated pentalinonsterol.
This test compound caused dose-dependent changes in the
content of fatty acid lipids in L. donovani promastigotes.
Pentalinonsterol shows potential for development as a therapy
for the treatment of visceral leishmaniasis.
Intellectual property protection through The Ohio State
University has been obtained.

(Gupta et al., ACS Inf. Dis., 2015, under revision)

INVESTIGATORS INVOLVED IN THE

ANTILEISMANIAL PROJECT ON PENTALINON ANDREUXII
FROM MEXICO AND FUNDING SOURCES
Phytochemical Studies The
Ohio State University
Prof. A. Douglas Kinghorn
Dr. Li Pan
Dr. Ben Naman

Synthetic Studies The Ohio

State University
Prof. James R. Fuchs
Dr. Dalia Abdelhamid

Biological
Testing
and
Mechanistic Studies The
Ohio State University
Prof. Abhay R. Satoskar
Prof. K.M. Ainslie
Prof. N.L. Pari Parinandi
Dr. Claudio Lezama-Dvila
Dr. Gaurav Gupta

Endophytic Fungal Cultivation

Studies Mycosynthetix, Inc.,
Hillsborough, NC

Collaborator (Taxonomy and

Supply of Plant Material
Universidad Autnoma de
Campeche, Mexico

Dr. Cedric J. Pearce

Dr. Anglica P. Isaac-Mrquez

Funding by grant RC-4 AI092624 was obtained from NIAID, NIH (A.R.
Satoskar, PI, A.D. Kinghorn, Co-PI; 2010-2013). Earlier support was obtained
from NIH grants R21 AI076309, R21 AT04160, and R56 AI090803 (A.R.
Satoskar, PI). Current funding is from the U.S. Army Medical Research
(W81XWH-14-2-0168; A.R. Satoskar, P.I.; 2014-2017)

SUMMARY AND CONCLUSIONS

Tropical plants are more biodiverse than
temperate plants, and thus hold the potential of
offering greater chemical diversity for natural
products drug discovery.
Plant collections to access plant genetic material
for drug discovery must cover the source country
in terms of benefit-sharing agreements.
Efforts to harness plant compounds as potential
cancer chemotherapeutic and antileishmanial
agents from an academic perspective require a
collaborative multidisciplinary approach with
open and frequent communications.

COLLEGE OF PHARMACY AND

OSU COMPREHENSIVE CANCER
CENTER